MICROMASS Q-Tof micro MASS SPECTROMETER

Transcription

1 Proudly serving laboratories worldwide since 1979 CALL for Refurbished & Certified Lab Equipment MICROMASS Q-Tof micro MASS SPECTROMETER QTOF Micro Performance Specifications Time of Flight Mass Resolution, Positive Ion 5000 (FWHM) on (M+H)+ ion from Leucine Enkephalin. Time of Flight Mass Resolution, Negative Ion 5000 (FWHM) on (M-H)- ion from Raffinose. Full Scan MS Sensitivity, Positive Ion The signal height obtained from a sample consumption of 200 fmol of horse heart Myoglobin (16952 Da) will be greater than 166 ion counts on the most intense peak in the charge state envelope. A solution of 200 fmol/µl horse heart Myoglobin (in 50/50 acetonitrile/water + 0.2% formic acid) will be introduced at a flow rate of 5 µl/min. Full Scan MS Sensitivity, Negative Ion The signal height obtained from the sample consumption of 1 ng of raffinose will be greater than 200 counts on the (M-H)- peak at m/z 503. This will correspond to a signal to noise ratio of greater than 200:1 (after a 1x3 smooth). The instrument will be tuned at 5000 resolution (as demonstrated in specification 2a) and a solution of 5 ng/l in 50/50 acetonitrile/water (no additives) will be introduced at 10 µl/min. Full Scan MS/MS Sensitivity, Positive Ion The signal height obtained from a consumption of 20 fmol of [Glu1] -Fibrinopeptide B (1569 Da) will be greater than 6 counts on the most intense yî sequence ion from the MS/MS spectrum of the doubly charged precursor ion. This will correspond to a signal to noise ratio of greater than 30:1 (after a 3x9 smooth) on the most intense yî sequence ion.

2 A solution of 100 fmol/µl will be introduced at a flow rate of 5 µl/min. Mass Measurement Accuracy (with internal reference) The RMS error between the measured and the accepted masses of peaks which have sufficient intensity and are free from interference from other masses, over the range from Daltons, will be less than 5 ppm. One suitable peak of known mass will be used as an internal reference. The instrument will be tuned at 5000 resolution as demonstrated in specification 1a. ELECTROSPRAY OPTION NANOFLOW Full Scan MS/MS Sensitivity, Positive Ion The signal to noise from a consumption of 2 fmol of [Glu1]-Fibrinopeptide B (1569 Da) will be greater than 30:1 (after a 3x9 smooth) on the most intense yî sequence ion from the MS/MS spectrum of the doubly charged precursor ion. A solution of 500 fmol/l concentration in MeOH/H2O + 0.2% formic acid solution and with glass micropipettes with 1 or 2 µm tips will be used. The integration period per spectrum will be about 5 sec and data will be summed over a period appropriate for the required consumption of sample. ELECTROSPRAY OPTION TRANSFORM SOFTWARE (Software for the determination of molecular weight from a spectrum containing a series of multiply charged ions on a m/z scale by a transform of the data to a true mass scale.) Mass Measurement Accuracy (no internal reference) The mean measured mass of transformed data shall be Da and the standard deviation (σ) of the mean <0.5 Da. The transform data will be created from five repeat analyses of the globin from normal human haemoglobin. Mass calibration to be performed using the multiply charged α globin peaks from a separate analysis. The raw data should be transformed over the range 15,000-16,000 Da and smoothed appropriately. The instrument will be tuned at 5000 resolution. It is recommended that a solution containing 10 pmol/µl of each globin in 50/50 acetonitrile/water + 0.2% formic acid is used.

3 TO IDENTIFY COMPONENTS IN YOUR SAMPLE, DO YOU NEED TO: Determine elemental composition? Elucidate structural characteristics? De novo sequence peptides with the highest confidence? Eliminate false positive protein identifications? Identify components in complex matrices? Distinguish between nominally isobaric amino acids? COMBINED ACCURACY AND SENSITIVITY Whether you want to identify and characterize proteins or perform structural elucidation of small molecules, exact mass measurement is as important as having the sensitivity to detect them. The Waters Micromass Q-Tof micro Mass Spectrometer enables you to combine sensitive LC/MS/MS with simple, automated exact mass measurement, increasing confidence in your results. The high quality data delivered by the Q-Tof micro can provide information on elemental composition, structural characteristics and excellent specificity for identifying your compounds in complex matrices. This benchtop, total solution integrates the Micromass Q-Tof micro Mass Spectrometer with NanoLockSpray and the Waters CapLC Pump and Autosampler using MassLynx 4.0 Software. WHY EXACT MASS? So, do you really know the mass of your compound? Are you confident that you ve identified the correct component? As with every analytical tool, the more accurately you make your measurement the more confidence you'll have in your results. Exact mass measurement enables you to determine the elemental composition of compounds with greater confidence. When you detect a drug metabolite at m/z in an in vivo sample, are you sure which bio-transformation has taken place? When you detect a peptide at m/z, can you be sure which posttranslational modification is present? With exact mass measurement you can:

4 Determine elemental composition Distinguish nominally isobaric components Improve specificity of detection for MS/MS analyses Improve data processing speed and specificity The inherent high resolution and mass accuracy of an orthogonal time-of-flight (oa-tof) mass spectrometer minimizes co-eluting and matrix interferences, enabling components of interest to be resolved. Automated exact mass measurements can then be performed "on the fly". The resolution of the Q-Tof micro is shown relative to the typical unit resolution used for full scan LC/MS analyses on a quadrupole or ion trap mass analyzer. The higher resolution of the ToF analyzeracross the full m/z range, in conjunction with exact mass measurement, enables unknowns to be more easily identified without losing precious information due to scan speed restrictions. The Waters Micromass Q-Tof micro Mass Spectrometer The Micromass Q-Tof micro is a high resolution oa-tof mass spectrometer that enables automated exact mass measurements in an easy-to-use benchtop instrument. The instrument also features a quadrupole mass filter and collision cell for MS/MS analyses. This powerful combination delivers simple exact mass measurement of fragment ions to yield increased confidence in structural elucidation and databank search results.

5 Why exact mass MS/MS? Exact mass measurement becomes an increasingly powerful tool for determining elemental composition at lower m/z values, and is therefore particularly useful for identification of fragment ions in MS/MS. With the increased accuracy of exact mass measurement, peptide sequencing and the elucidation of small molecule structures is significantly enhanced. Data processing tools for automated interpretation rely heavily on the specificity of exact mass measurements. With exact mass MS/MS you can: Eliminate false positive protein ID's apparent in nominal mass data De novo sequence peptides with the highest level of confidence Identify elemental composition of structural fragments Gain excellent specificity for quantitative analyses

6 Exact mass MS measurement enables compounds to be more easily identified. Confirming a measured m/z to within 5 ppm significantly increases confidence in assigning elemental composition. De novo sequencing of peptides is significantly enhanced through the specificity of exact mass MS/MS. The table demonstrates how greater confidence in sequence assignment is achieved with improved mass accuracy. Results are shown for a digest of Bovine Serum Albumin using ProteinLynx Global SERVER 2.0 Software.

7 A powerful technology platform The Q-Tof micro provides a powerful platform for MS and MS/MS analyses, upon which total system solutions may be designed to maximize data quality for your application. The Q-Tof micro Mass Spectrometer offers: A small footprint powerful Q-Tof technology on your benchtop ZSpray dual orthogonal source technology for unsurpassed sensitivity and ruggedness High resolution 5000 FWHM for resolving nominally isobaric ions Exact mass measurement sub 5 ppm RMS in both MS and MS/MS modes Extended linear dynamic range for quantitative analysis oa-tof analyzer wide acquisition range of up to 20,000 m/z Data Directed Analysis (DDA ) intelligent detection of compounds of interest for automated switching from MS to MS/MS Precursor Ion Discovery patented acquisition techniques for class-specific analyses (i.e. detection of specific structural motifs) using exact neutral loss or product ion detection Variable Flow Chromatography with the Waters CapLC System for enhanced MS/MS data quality Ease of use wizard-driven system configuration and method editing for simplified operation

8 Exact mass MS/MS is achieved automatically using the Q-Tof s LockSpray dual electrospray source. The LockSpray dual electrospray source enables automated exact mass measurement with an infused internal lockmass from a second sprayer, eliminating the need for T-plumbing and potential ionization interference between analytes and standard. The NanoLockSpray dual electrospray source is optimized for exact mass data acquisition with nanoflow LC/MS/MS and is ideal for proteomics studies or low level metabolite identification. Systematic productivity Whether you are characterizing peptides and proteins, or identifying small molecules, such as metabolites or impurities, in complex matrices, Waters Mass Spectrometry Systems incorporating the Q-Tof micro maximize performance for your application. The fully integrated systems are controlled from a single data system, streamlining your analysis while enabling advanced experiments to be easily performed.

9 The Waters Mass Spectrometry System for Metabolite Identification integrates the Waters 1525µ Binary HPLC Pump, 2777 Sample Manager, 2996 Photodiode Array Detector, Q-Tof micro with LockSpray, all controlled by MassLynx 4.0 Software with the MetaboLynx Application Manager. Protein Characterization Comprehensive chromatographic capabilities for complex mixture separation Exact mass MS and MS/MS for the highest confidence in databank searching and de novo sequencing Exact mass precursor ion discovery for confident detection and characterization of modified peptides Automated protein identification and characterization with MS-WorkFlow ProteinLynx Global SERVER 2.0 Determination of intact protein complexes with Maximum Entropy processing METABOLITE/IMPURITY IDENTIFICATION Sensitive full spectral acquisition for detection of predicted and unexpected components Exact mass MS and MS/MS for structural comparison with parent drug MetaboLynx Application Manager for intelligent exclusion of endogenous components and automated processing of exact mass MS and MS/MS from a single analysis METABONOMICS Sensitive full scan MS for detection of low level endogenous metabolites Exact mass MS and MS/MS for accurate identification High quality liquid chromatography for efficient separation and high retention time reproducibility High quality LC/MS data for efficient statistical analysis of the samples Natural Product Identification Sensitive full scan MS for detection of multiple components Exact mass MS and MS/MS for accurate identification Integrated high throughput LC with environmental control for samples Automated exact mass data processing Intelligent LC/MS/MS With both throughput and sample volume restrictions on many analyses, it is increasingly important to maximize the amount of real information that can be extracted from a single experiment. Integrating your Waters chromatography and MS instrumentation into an intelligent mass spectrometry system with

10 MassLynx Software enables maximum sensitivity and selectivity for your components of interest. Intelligent Data Directed Analysis (DDA) DDA enables intelligent MS and MS/MS analyses to be performed automatically, maximizing the amount of real information acquired on components of interest. Using the easy-to-use DDA Wizard, your analysis can be targeted for detection of analytes based on charge state recognition, exact mass and peak intensity. Exclude lists can also be employed to avoid redundant analyses. Detection in the MS Survey mode triggers a switch to MS/MS where exact mass data are quickly acquired on structural fragments. Advanced Application Managers such as ProteinLynx Global SERVER 2.0 and MetaboLynx have been designed for rapid processing of DDA data, increasing the efficiency of both your analysis and results reporting. Precursor ion discovery (PID) DDA can be used to automate precursor ion discovery. The Q-Tof micro employs a novel acquisition mode that enables highly selective detection of an exact neutral loss (e.g. from phosphopeptides) or product ion (e.g. glycans) to trigger MS/MS on components of the related precursor ion. By cycling collision energy between high and low modes in MS survey, the software can very accurately detect targeted product ions or neutral losses. Identification of these triggers selection of the precursor for true MS/MS. Intelligent MS and MS/MS data acquisition in a single run using Data Directed Analysis (DDA).

11 Exact neutral loss of the glucuronide conjugate ( Da) from a Phase II metabolite is detected in MS mode by cycling collision energy. The identified precursor is subsequently selected for MS/MS analysis. Advanced chromatography integration The integration of Waters LC and MS instruments through a single data system enables a simplification of operation, while also creating a platform for more advanced experiments. By using exact mass MS detection, analyses can be automatically directed in real time based on pre-defined criteria. Maximizing efficiency and sensitivity Capillary LC, utilizing the Waters CapLC System, benefits from very efficient chromatographic peaks, enabling complex mixtures to be resolved and more specific MS and MS/MS data to be obtained. An additional benefit is the inherent increase in sensitivity provided by the CapLC due to concentration effects, yielding much more sensitive MS detection. Variable flow chromatography (VFC) Variable flow chromatography can automatically reduce the flow rate of the CapLC upon detection of a specified ion by the Q-Tof micro Mass Spectrometer. This can be of particular use in a precursor ion discovery experiment, increasing the length of MS/MS acquisition time available for the analysis of targeted precursor ions.

12 Phosphopeptide analysis by PID on the Q-Tof micro. VFC allows MS/MS data to be acquired from the targeted ion for 3 minutes (compared to only 30 seconds at standard flow rate), resulting in increased structural information and a >10x improvement in signal-to-noise. 2D LC Multi-dimensional chromatography can also be driven through the Waters Mass Spectrometry System that features the CapLC and Q-Tof micro. The ability to resolve components across different column chemistries is particularly useful for complex mixture analysis. The Stream Select Module can be easily configured to include an additional ion exchange column, enabling complex peptide mixtures to be resolved based on charge prior to their separation on a reversed phase capillary analytical column.

13 2D LC configured for the CapLC and Q-Tof micro using the Stream Select Module. Chromatography-based columns and reagents Speed and sensitivity are optimized by choosing the correct column and extraction products. Symmetry and Symmetry300 Columns To meet your LC/MS method development needs, Symmetry columns provide the highest standard of "column-to-column" reproducibility with unmatched peak symmetry for maximum sensitivity and accurate quantification. Atlantis Columns Atlantis columns are a difunctionally bonded, silica-based C18 offering that combines superior polar compound retention with exceptional peak shape and full LC/MS compatibility. XTerra MS Columns XTerra MS columns, based on a combination of our most inert particle and trifunctional reversed-phase bonding technology, are ideally suited for mass spectrometry applications. Capillary, NanoEase and PicoFrit nano, and Opti-Pak Trapping Columns A complete offering of capillary, nano and trapping column technologies that can be effectively utilized to capture and concentrate peptides in either 1D or 2D LC or LC/MS applications. RapiGest SF An enabling agent that dramatically improves "in-solution" protein digestions in terms of speed and peptide recovery with no interference in LC or LC/MS applications. OVERVIEW

14 Figure 1-1 The Micromass Q-tof micro Instrument Description The Q-Tof micro hybrid quadrupole time of flight mass spectrometer is available with electrospray ionization (ESI) and atmospheric pressure chemical ionization (APcI). Q-Tof micro utilizes a high performance, research grade quadrupole mass analyzer, incorporating a prefilter assembly to protect the main analyzer from contaminating deposits, and an orthogonal acceleration time of flight (TOF) mass spectrometer. A hexapole collision cell, between the two mass analyzers, can be used to induce fragmentation to assist in structural investigations. Ions emerging from the second mass analyzer are detected by the microchannel plate detector and ion counting system. A PC computer runs the MassLynx NT software system to control Q-Tof micro, and to acquire and process data. Ionization Techniques Using the Micromass Z-spray atmospheric pressure ionization (API) source, two techniques are available. Atmospheric Pressure Chemical Ionization Atmospheric pressure chemical ionization (APcI) generally produces protonated or deprotonated molecular ions from the sample via a proton transfer (positive ions) or proton abstraction (negative ions) mechanism. The sample is vapourised in a heated nebulizer before emerging into a plasma consisting of solvent ions formed within the atmospheric source by a corona discharge. Proton transfer or abstraction then takes place between the solvent ions and the sample. Eluent flows up to 2 milliliters/minute can be accommodated without splitting the flow. Electrospray Electrospray ionization (ESI) takes place as a result of imparting a strong electrical charge to the eluent as it emerges from the nebulizer. An aerosol of charged droplets emerges from the nebulizer. These undergo a reduction in size by solvent evaporation until they have attained a sufficient charge density to allow sample ions to be ejected from the surface of the droplet ( ion evaporation ). A characteristic of ESI spectra is that ions may be singly or multiply charged. Since the mass spectrometer filters ions according to their mass-to-charge ratio ( ), compounds of high molecular weight can be determined if multiply charged ions are formed. Eluent flows up to 1 ml/min can be accommodated although it is often preferable with electrospray ionization to split the flow such that 5-50 µl/min of eluent enters the mass spectrometer. Nanoflow Electrospray The optional nanoflow interface allows electrospray ionization to be performed in the flow rate range 5 to 1000 nanolitres per minute. For a given sample concentration, the ion currents observed in nanoflow are comparable to those seen in normal flow rate electrospray. Great sensitivity gains are therefore observed when similar scan parameters are used, due to the great reductions in sample consumption.

15 Ion Optics Q-Tof micro Ion Optics The principal components of the ion optical system are shown in Figure 1-2. Ions generated in the Z-spray source are transferred to the quadrupole analyzer MS1 via the independently pumped RF lens. After leaving the quadrupole analyzer the ions flow into the orthogonal time of flight analyzer MS2. The ion beam is focused into the pusher by the acceleration, focus, steer and tube lenses. The pusher then pulses a section of the beam towards the reflectron, which then reflects ions back to the detector. As ions travel from the pusher to the detector they are separated in mass according to their flight times, with ions of the highest mass to charge ratio ( ) arriving later.

16 The pusher may be operated at repetition frequencies of up to 30 khz, resulting in a full spectrum being recorded by the detector every 33 microseconds. Each spectrum is summed in the histogram memory of the time to digital converter until the histogrammed spectrum is transferred to the host PC. If the user has requested an acquisition rate of 1 spectrum/second, each spectrum viewed on the host PC will be the result of summing up to 30,000 individual spectra recorded at the detector. Unlike scanning instruments, the TOF performs parallel detection of all masses within the spectrum at very high sensitivity and acquisition rates. This characteristic is of particular advantage when the instrument is coupled to fast chromatography, since each spectrum is representative of the sample composition at that point in time, irrespective of how rapidly the sample composition is changing. Mechanical Components Mechanical Components - Internal View.

17 The main internal mechanical components of the instrument are: The source housing, containing the RF (hexapole) lens. The MS1 analyzer housing, containing the quadrupole analyzer, hexapole collision cell and hexapole transfer lens The TOF analyzer housing, containing the pusher, detector and reflectron assemblies. One 250 litre/second turbomolecular pump, plus one split-flow turbomolecular pump. Two active inverted magnetron (Penning) gauges and two Pirani gauges. Electrical Components The main electronics unit is located in the lower rear section of the instrument. This contains: High voltage power supplies. These supply the probe or corona, reflectron, TOF flight tube and lens circuits. Low voltage power supplies. These supply the PCBs, high voltage supplies and turbomolecular pumps. Main PCBs. For communications, lenses and quadrupole control.

18 Rear View of Instrument

19 The Vacuum System Figure 1-5 Q-Tof micro Vacuum System Fine Pumping Q-Tof micro is equipped with three water cooled turbomolecular pumps, providing independent fine pumping of the source hexapole, quadrupole and TOF analyzers. Details of the operation and maintenance of the pumps can be found in the manufacturer s manuals provided. Rotary Pumping Source pumping and turbomolecular pump backing is by a direct drive rotary pump. The rotary pump is situated at the front of the instrument. Details of the operation and maintenance of the pump can be found in the manufacturer s manual provided. Pressure Measurement The backing pressure is monitored by an active Pirani gauge. The analyzer and TOF pressures are monitored by active inverted magnetron (Penning) gauges. These gauges act as vacuum switches, switching the instrument out of Operate mode if the pressure is too high. Pressure readings may be displayed on the MassLynx NT tune page. The analyzer Penning gauge only comes on when the vacuum display window is open. At other times the gauge is off. The analyzer Pirani gauge is used when the diaply is off, though no pressures are shown. Vacuum Protection

20 The vacuum system is fully interlocked to provide adequate protection in the event of: a fault in the vacuum system. a failure of the power supply. a failure of the water supply. a vacuum leak. 1.5 Front Panel Connections Front Panel Desolvation Gas and Probe Nebulizer Gas The PTFE gas lines for the Desolvation Gas and probe Nebulizer Gas are connected to the front of the instrument using threaded metal fittings. Cone Gas is connected internally. High Voltage The electrical connection for the ESI capillary or the APcI corona discharge pin is via the coaxial high voltage connector. This socket is labeled Capillary / Corona. Heaters The electrical connection for the APcI probe or the ESI desolvation heater is via the

21 multi-way connector labeled Probes. This is removed from the front panel by pulling on the metal sleeve of the plug. Both the electrospray desolvation heater and the APcI probe heater use this connector. The power for the source block heater is permanently connected. As a consequence, the source block assembly is usually very hot, and should not be touched. Front Panel Controls and Indicators Status Display The display on the front panel of the instrument consists of two 3-colour light emitting diodes (LEDs). The display generated by the Pump LED is dependent on the vacuum status of the instrument. The Operate LED depends on both the vacuum status and whether the operate mode has been selected from the Data System. Further information is included in Automatic Pumping and Vacuum Protection (see Routine Procedures). Divert / Injection Valve The divert / injection valve may be used in several ways depending on the plumbing arrangement: As an injection valve, with the needle port and sample loop fitted. As a divert valve, to switch the flow of solvent during a LC run. As a switching valve to switch, for example, between a LC system and a syringe pump containing calibrant. This valve is pneumatically operated, using the same nitrogen supply as the rest of the instrument. The two switches marked Load and Inject enable the user to control the valve when making loop injections at the instrument. 1 Rear Panel Connections

22 Water Water is used to cool the turbomolecular pumps. Nitrogen Gas In The nitrogen supply (100 psi, 7 bar) should be connected to the Nitrogen Gas In push-in connector using 6mm PTFE tubing. If necessary this tubing can be connected to 1/4 inch tubing using standard 1/4 inch fittings. Caution: Use only PTFE tubing or clean metal tubing to connect between the nitrogen supply and the instrument. The use of other types of plastic tubing will result in chemical contamination of the source. Exhausts The exhaust from the rotary pump should be vented to atmosphere outside the laboratory. The gas exhaust, which also contains solvent vapours, should be vented via a separate fume hood, industrial vent or cold trap. The gas exhaust should be connected using 10mm plastic tubing connected to the push-in fitting. Caution: Do not connect these two exhaust lines together as, in the event of an instrument failure, rotary pump exhaust could be admitted into the source chamber producing severe contamination.

23 Supply Inlet The mains power cord should be wired to a 230V mains outlet using a suitable plug, or to a transformer. For plugs with an integral fuse, the fuse should be rated at 13 amps (UK only). Electronics This circuit breaker switches power to the electronics. In the event of the instrument drawing more than the rated current, the circuit breaker will trip. Rotary Pump This circuit breaker switches power to the rotary and turbomolecular pumps. In the event of the pumps drawing more than the rated current, it will trip. Event Out Four outputs, Out 1 to Out 4 (Figure 1-8), are provided to allow various peripherals to be connected to the instrument. Switches S1 to S4 allow each output to be set to be either a contact closure (upper position) or a voltage output (lower position). Out 1 and Out 2, when set to voltage output, each have an output of 5 volts. The voltage output of both Out 3 and Out 4 is 24 volts. During a sample run an event output may be configured to close between acquisitions and is used typically to enable an external device to inject the next sample. Contact Closure In In 1 and In 2 inputs are provided ((Figure 1-8) to allow an external device to start sample acquisition once the device has performed its function (typically sample injection). Analog Channels Four analog channel (Figure 1-8) inputs are available, for acquiring simultaneous data such as a UV detector output. The input differential voltage must not exceed one volt.

24 Rear Panel Outputs MassLynx Data System A PC computer runs the MassLynx NT software system to control Q-Tof micro, and to acquire and manipulate data from it. A high resolution colour monitor is also supplied. Interaction with MassLynx NT is via the mouse and keyboard using menu-driven commands. Printing, file management and other routine procedures are performed using the appropriate Windows NT modules. Software The following software packages are supplied with Q-Tof micro: MassLynx NT. DataBridge, a utility to convert other format data files into MassLynx format. Microsoft Windows NT/2000/XP graphical environment. Mouse configuration. A range of optional software modules for different applications is also available. The MassLynx NT User s Guide describes the many facilities of the Micromass software.

25 INSTRUMENT SPECIFICATIONS To enable the instrument to give its best performance the recommended environmental conditions and power and water supplies are outlined below. Preparation of the laboratory in advance will assist us in efficiently installing the instrument. ROOM LOCATION Dimensions The Micromass Q-Tof micro MS is 180 mm wide by 635 mm deep by 1163 mm long and weighs 200 kg. It is mounted on 6 supporting feet. An external Edwards EM28 requires an additional 650 mm by 200 mm of floor space. In the event that the unit needs to be lifted, the side panels should be removed and suitable lifting bands passed beneath the bottom of the frame. These should be attached to a suitable device (hoist etc.) to raise the analyzer unit in a safe and controlled way. Only trained personnel with the correct equipment should carry this out. A separate table 1200 mm by 730 mm is supplied for the computer terminal. Doorways through which the instrument is to be transported should be a minimum of 820 mm wide. In the laboratory a minimum clearance of 500 mm should be allowed all round the bench for service access, apart from at the rear of the instrument where 900 mm is required. Note: The instrument should not be placed close to heavy machinery (compressors, generators etc.) which give excessive floor vibration. ENVIRONMENT It is recommended that the instrument be sited in an air conditioned laboratory, in a draught free position and away from excessive amounts of dust. GENERAL Altitude: upto 2000m Pollution degree 1 in accordance with IEC 664 Rotary Pump 150 to 400C Instrument The maximum ambient laboratory temperature should not exceed 300C, optimum temperature lies in the range C. Short term (1.5 hour) variations should be no more than 20C.

26 The relative humidity should not exceed 70%. Heat dissipated into the laboratory from the instrument is about 1.2 kw. The instrument conforms to IEC , Pollution Degree 1, and Installation Category II. MAGNETIC FIELDS OA-Tof instruments are relatively tolerant to stray magnetic fields. We would advise an upper limit of 10 Gauss for both AC and DC components of magnetic field measured at the mass spectrometer. RADIO EMISSIONS The instrument should not be placed within a RF field greater than 0.2 V/metre. This approximates to a 1W hand held unity gain transmitter at a distance of 10 m. Possible sources of RF emission include RF linked alarm systems or LANs, portable telephones and hand held transmitters. WATER SUPPLIES The heat dissipated into the cooling water is about 400 Watts. The water flow required to dissipate the heat generated by the turbo pumps is 35 L/hour for an inlet temperature of about 200 C or 23 L/hour at an inlet temperature of about 150 C assuming an outlet temperature of about 300 C. The water may be supplied by a recirculating chiller with the following characteristics: Heat dissipation into system: 400 W Temperature stability: +/- 20C Minimum reservoir volume: 5 L Minimum supply pressure:10 psi (outlet at atmospheric pressure). Maximum supply pressure: 60 psi Minimum flow rate at 150C: 0.4 L/min The above assume that the outlet water temperature will be no more than 300C. Inlet water temperatures below 150C are not recommended since excessive condensation may form on exposed pipework. Alternatively, when there is a cooled water supply available it may be used either directly through the instrument or indirectly via a water-to-water heat exchanger. In this latter case the chilled water supply to the heat exchanger must be at least 100C below the required inlet temperature for the instrument. One inlet and one outlet are required for the instrument. Reinforced 10mm (3/8 inch) flexible hose is preferred. To prevent blockage of the water pipes suitable in-line filters will be required to remove particulate matter from town water supplies when these are used.

27 Operation above 2000 metres altitude may adversely affect the cooling of the system. POWER REQUIREMENTS The instrument requires a single phase Hz, 230 V nominal power supply rated at 13 A (UK) or 15 A (Europe). An additional single phase Hz, 230 V or 115 V nominal power supply rated at 5A (UK) is required to run the embedded PC. In the USA and Canada a single-phase Hz supply at 230 V phase to neutral fused and rated at 15 A is required. Alternatively two phases of a Hz 208 V phase to phase, 3 phase supply, rated and fused at 15 A may be used. It is mandatory that no other apparatus is connected to this supply. Circuit breakers are an acceptable alternative to fuses. The supply should be terminated in the laboratory no more than 2 m from the instrument with either a wall mounted isolator or socket and plug to be fitted to the instrument. Other supply voltages can be accommodated using a transformer to change the primar supply voltage to 230 V. Advance notice is required and Micromass should be contacted. On single-phase supplies the power supply should ensure that the line and neutral wires cannot be transposed. On pump start-up currents of up to 30 A may be drawn for several seconds. Time delay fuses and breakers are recommended to prevent nuisance tripping. A safety earth (ground) correctly rated must be provided in all cases. Data system components, chromatographs, syringe pumps etc. should be connected directly to laboratory power outlets (no ancillary outlets are provided on the instrument). A residual current device (RCD) is recommended for additional protection. In the case of instruments fitted with a transformer the RCD should be fitted in the supply side of the transformer. Supply brownout should not fall to less than half main voltage for greater than 20 msec duration. GASES AND REGULATORS Nitrogen The instrument requires oil free dry nitrogen regulated at 7 bar (100 psi) minimum outlet pressure to provide nebulising and drying gas to the instrument. During API operation typical usage of nitrogen is about 400 L/hour, but under high flow rate conditions (Megaflow/APcI) this may increase to 650 L/hour. This equates, approximately, to the consumption of a large cylinder of compressed nitrogen each day and it may be preferred to use a liquid nitrogen dewar which may last several weeks. Collision Gases Typically Argon is used as the collision gas for CID experiments. This should be 99.9% pure, regulated at no more than 50 psi. Connection is via 1/8 inch OD stainless steel or copper tubing (NOT SUPPLIED).

28 EXHAUST OUTLETS Rotary Pump Outlet The rotary pump exhaust outlet must be vented to the atmosphere external to the laboratory clear from any air intakes for air conditioning systems. A 12 mm (1/2 inch) hose connection is required. If the length of exhaust exceeds 4 m then the internal diameter of the pipe should be increased to 48 mm (2 inch) for the excess distance. Nitrogen Outlet Severe damage to the instrument will result if the electrospray/apci exhaust is connected to the rotary pump exhaust line. This will occur when the nitrogen supply is off and rotary pump oil vapour will migrate via the source exhaust to the ion source and then through the sampling orifice into the quadrupole and gas cell assembly. A separate exhaust for the ion source gas (nitrogen) must be provided to the atmosphere external to the laboratory clear from any air intakes for air conditioning systems. A (6mm OD) hose connection is required. If the length of exhaust exceeds 3 m then the internal diameter of the pipe should be increased to 12 mm (1/2 inch) for the excess distance. Proudly serving laboratories worldwide since 1979 CALL for Refurbished & Certified Lab Equipment